Library Preparation for Next Generation Sequencing

PureGenome™ High Efficiency NGS Library Preparation Reagents

PureGenome™ High Efficiency Next Generation Sequencing (NGS) Library Preparation is a uniquely optimized system of three library preparation reagent sets, collectively requiring less hands-on time than similar existing protocols.

The ease of use two-step procedure is followed by library amplification using Merck’s ultra-high fidelity KOD Hot Start DNA Polymerase Master mix. This unique polymerase amplifies DNA with high processivity in highly TA- or GC-rich regions. The combined efficiency of library construction and accuracy of amplification enables maximum library yields from lower input DNA-with minimal bias for better results.

Consolidated NGS Library Preparation Workflow: Two Steps in Under Two Hours

Maximum Coverage with Minimum Bias

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Two micrograms of sheared E. coligenomic DNA was used to generate an Illumina/Solexa pair-end compatible library using the PureGenome™ High Efficiency NGS library preparation reagent sets and competitor kits. Libraries were sequenced using a Illumina MiSeq® instrument. In our process, nine cycles of PCR (using KOD Hot Start DNA Polymerase) were used to enrich the library. The library (10 pM) bioanalyzer profile is shown (A). The PureGenome™ NGS Library Reagents resulted in an average of 150x genome coverage (B) without GC bias (C). The accuracy of sequencing demonstrates the high quality of NGS libraries constructed with PureGenome™ High Efficiency NGS Library Preparation reagents (D).